WO2020097460A1 - Nanoparticules de vegf auto-assemblées - Google Patents

Nanoparticules de vegf auto-assemblées Download PDF

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Publication number
WO2020097460A1
WO2020097460A1 PCT/US2019/060466 US2019060466W WO2020097460A1 WO 2020097460 A1 WO2020097460 A1 WO 2020097460A1 US 2019060466 W US2019060466 W US 2019060466W WO 2020097460 A1 WO2020097460 A1 WO 2020097460A1
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Prior art keywords
peptide
segment
nanofiber
amphiphile
peptide amphiphile
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Puneet OPAL
Samuel I. Stupp
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Northwestern University
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Northwestern University
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/52—Cytokines; Lymphokines; Interferons
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/18—Growth factors; Growth regulators
    • A61K38/1858—Platelet-derived growth factor [PDGF]
    • A61K38/1866—Vascular endothelial growth factor [VEGF]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery

Definitions

  • PAs self-assembling peptide amphiphiles
  • PAs comprising a bioactive vascular endothelial growth factor (VEGF) peptide, nanofibers displaying VEGF PAs, and methods of treatment or prevention of the polyglutamine disease Spinocerebellar Ataxia Type 1 (SCA1) and other neurodegenerative diseases therewith.
  • VEGF peptide delivery platform is provided in which the mechanical properties of the nanofiber material are tunable by altering the ratio of bioactive PA to structural PAs (e.g., acidic or basic PAs lacking a bioactive epitope).
  • Alzheimer’s, Parkinson’s, amyotrophic lateral sclerosis, Huntington’s disease and other polyglutaminopathies are clinically and etiologically heterogeneous. Yet it is increasingly apparent that these diseases, long considered strictly in terms of neuronal dysfunction, also disrupt the close coupling of neural activity with blood flow (Iadecola, 2017; herein incorporated by reference in their entireties).
  • VEGF vascular endothelial growth factor
  • VEGF may also be therapeutic in
  • Parkinson’s disease (Caballero et al, 2017; Zou et al, 2017; herein incorporated by reference in their entireties), Alzheimer’s disease (Echeverria et al, 2017; herein incorporated by reference in its entirety), and amyotrophic lateral sclerosis (Wang et al, 2016; herein incorporated by reference in its entirety).
  • VEGF mimetic peptide A completely synthetic VEGF mimetic peptide was developed to overcome these issues, in which a fifteen-amino acid VEGF sequence (KLTWQELYQLKYKGI (SEQ ID NO: 1) is covalently linked to an amphiphilic peptide (Webber et al, 2011; herein incorporated by reference in its entirety). These amino acids mimic VEGF residues 17 through 25 in their ability to bind and activate VEGF receptors (D'Andrea et al, 2005;
  • VEGF-PA VEGF-mimetic peptide amphiphile
  • Nano-VEGF had never been tested in the brain.
  • PAs self-assembling peptide amphiphiles
  • PAs comprising a bioactive vascular endothelial growth factor (VEGF) peptide, nanofibers displaying VEGF PAs, and methods of treatment or prevention of the polyglutamine disease Spinocerebellar Ataxia Type 1 (SCA1) and other neurodegenerative diseases therewith.
  • VEGF peptide delivery platform is provided in which the mechanical properties of the nanofiber material are tunable by altering the ratio of bioactive PA to structural PAs (e.g., acidic or basic PAs lacking a bioactive epitope).
  • peptide amphiphile nanofibers comprising: (a) a bioactive peptide amphiphile comprising: (i) a hydrophobic non-peptidic segment; (ii) a b-sheet-forming peptide segment; (iii) a charged peptide segment; and (iv) a VEGF peptide comprising at least 50% sequence identity with KLTWQELYQLKYKGI (SEQ ID NO: l); and (b) a charged peptide amphiphile comprising: (i) a hydrophobic non-peptidic segment;
  • the charged peptide amphiphile does not comprise a bioactive peptide.
  • the hydrophobic non-peptidic segment of the bioactive peptide amphiphile and the charged peptide amphiphile comprises an acyl chain.
  • the acyl chain comprises C6-C20. In some embodiments, the acyl chain comprises Cl 6.
  • the b-sheet-forming peptide segment of the bioactive peptide amphiphile and the charged peptide amphiphile comprises AAAVVV (SEQ ID NO:2) or AAVV (SEQ ID NO:3).
  • the charged peptide segment of the bioactive peptide amphiphile is an acidic peptide segment.
  • the acidic peptide segment of the bioactive peptide amphiphile comprises Glu (E) and/or Asp (D) residues.
  • the acidic peptide segment of the bioactive peptide amphiphile comprises is 2- 7 amino acids in length with 50% or more amino acids selected from Glu (E) and/or Asp (D) residues.
  • the acidic peptide segment of the bioactive peptide amphiphile comprises EEE.
  • the charged peptide segment of the bioactive peptide amphiphile is a basic peptide segment.
  • the basic peptide segment of the bioactive peptide amphiphile comprises one or more lysine (K), histidine (H), and/or arginine (R) residues.
  • the basic peptide segment of the bioactive peptide amphiphile comprises is 2-7 amino acids in length with 50% or more Lys (K) residues.
  • the basic peptide segment of the bioactive peptide amphiphile comprises KKK.
  • the charged peptide segment of the charged peptide amphiphile is an acidic peptide segment.
  • the acidic peptide segment of the charged peptide amphiphile comprises Glu (E) and/or Asp (D) residues. In some embodiments, the acidic peptide segment of the charged peptide amphiphile comprises is 2-7 amino acids in length with 50% or more amino acids selected from Glu (E) and/or Asp (D) residues. In some embodiments,
  • the acidic peptide segment of the charged peptide amphiphile comprises EEE.
  • the charged peptide segment of the charged peptide amphiphile is a basic peptide segment.
  • the basic peptide segment of the charged peptide amphiphile comprises one or more lysine (K), histidine (H), and/or arginine (R) residues.
  • the basic peptide segment of the charged peptide amphiphile comprises is 2-7 amino acids in length with 50% or more Lys (K) residues.
  • the basic peptide segment of the charged peptide amphiphile comprises KKK.
  • the VEGF peptide comprises at least 70% sequence identity with SEQ ID NO: 1. In some embodiments, the VEGF peptide comprises SEQ ID NO: 1.
  • the peptide amphiphile nanofiber comprises 5%-75% by mass bioactive peptide amphiphile and 25% to 95% by mass basic peptide amphiphile, and wherein the nanofiber forms a gel under basic conditions.
  • the peptide amphiphile nanofiber comprises 5%-75% (by mass) bioactive peptide amphiphile and 25% to 75% (by mass) basic peptide amphiphile, and wherein the nanofiber forms a gel under basic conditions.
  • the peptide amphiphile nanofiber comprises 75-99% (by mass) bioactive peptide amphiphile and 1% to 25% (by mass) basic peptide amphiphile, and wherein the nanofiber is a liquid under basic conditions. In some embodiments, 75%, 80%, 85%, 90%, 95%, 99%, or ranges therebetween bioactive peptide amphiphile. In some embodiments, 1%, 5%, 10%, 15%, 20%, 25%, or ranges therebetween basic peptide amphiphile.
  • the peptide amphiphile nanofiber comprises 1-20% (by mass) bioactive peptide amphiphile and 80-99% (by mass) acidic peptide amphiphile, and wherein the nanofiber forms a gel under acidic conditions. In some embodiments, 1%, 5%, 10%, 15%, 20%, or ranges therebetween bioactive peptide amphiphile. In some embodiments, 80%,
  • the peptide amphiphile nanofiber comprises 20-80% (by mass) bioactive peptide amphiphile and 20-80% (by mass) acidic peptide amphiphile, and wherein the nanofiber forms a gel under neutral conditions.
  • the peptide amphiphile nanofiber comprises 80-99% (by mass) bioactive peptide amphiphile and 1-20% (by mass) acidic peptide amphiphile, and wherein the nanofiber is a liquid under acidic conditions. In some embodiments, 80%, 85%, 90%, 95%, 99%, or ranges therebetween bioactive peptide amphiphile. In some embodiments, 1%, 5%, 10%, 15%, 20%, or ranges therebetween acidic peptide amphiphile.
  • a peptide amphiphile nanofiber comprising: (a) a bioactive peptide amphiphile comprising: (i) a hydrophobic non-peptidic segment comprising a C6-C20 acyl chain; (ii) a b-sheet-forming peptide segment comprising
  • AAAVVV (SEQ ID NO: 2) or AAVV (SEQ ID NO: 3);
  • a charged peptide segment wherein the charged peptide segment comprises: (A) an acidic peptide segment comprising EEE, EED, EDE, DEE, EDD, DED, DDE, or DDD; or (B) a basic peptide segment comprising 2-7 or more lysine (K), histidine (H), and/or arginine (R) residues; and (iv) a VEGF peptide comprising at least 50% sequence identity with KLTWQELYQLKYKGI (SEQ ID NO: 1); and (b) a charged peptide amphiphile comprising: (i) a hydrophobic non- peptidic segment comprising a C6-C20 acyl chain; (ii) a b-sheet-forming peptide segment comprising AAAVVV (SEQ ID NO: 2) or AAVV (SEQ ID NO: 3); and (i
  • a peptide amphiphile herein comprises a linker segment between the charged peptide segment and the bioactive peptide segment.
  • the linker segment comprises 1-3 glycine (G) residues.
  • a bioactive peptide amphiphile herein comprises with at least 70% sequence identity (e.g., 70%, 75%, 80%, 85%, 90%, 95%, or ranges therebetween) with one selected from one of C16-V3A3E3G- KLTWQELYQLKYKGI (SEQ ID NO: 8), C16- V3A3E4G- KLTW QELY QLKYKGI (SEQ ID NO: 9), C16-V2A2E2G- KLTWQELY QLKYKGI (SEQ ID NO: 10), C16-V2A2E4G- KLTW QELY QLKYKGI (SEQ ID NO: 11), C16-V2A2E4G4- KLTW QELY QLKYKGI (SEQ ID NO: 12), C16-A2G2E4G- KLTWQELY QLKYKGI (SEQ ID NO: 13), C16-VEVA2E4G- KLTWQEL
  • a bioactive peptide amphiphile selected from one of C16-V3A3E3G- KLTWQELYQLKYKGI (SEQ ID NO: 8), C16-V3A3E4G- KLTWQELYQLKYKGI (SEQ ID NO: 9), C16-V2A2E2G- KLTWQELY QLKYKGI (SEQ ID NO: 10), C16-V2A2E4G- KLTWQELYQLKYKGI (SEQ ID NO: 11), C16-V2A2E4G4- KLTWQELYQLKYKGI (SEQ ID NO: 12), C16-A2G2E4G- KLTWQELYQLKYKGI (SEQ ID NO: 13), C16-VEVA2E4G- KLTWQELYQLKYKGI (SEQ ID NO: 14), C16-V2A2K3G- KLTWQELYQLKYKGI (SEQ ID NO: 9
  • a neurologic condition comprising administering a pharmaceutical composition comprising a peptide amphiphile nanofiber described herein to a subject suffering from the neurologic condition.
  • the neurologic condition is a poly glutamine disease.
  • the poly glutamine disease Spinocerebellar Ataxia Type 1.
  • the pharmaceutical composition is administered parenterally.
  • the pharmaceutical composition is administered by intrathecal administration,
  • intracerebroventricular administration or intraparenchymal administration.
  • FIG. 1A-D Nano-VEGF delivery improves vascular physiology in SCA1 mice.
  • (C) Nano- VEGF and rVEGF treatment significantly increase dendritic length and improves morphology of Purkinje cells in SCA1 mice as assayed by calbindin staining (scale bar lOOpm). This is quantified in D.
  • FIG. 2A-H Nano-VEGF significantly rescues cerebellar motor phenotype in late symptomatic SCA1 mice with effects comparable to or better than recombinant VEGF
  • C VEGF treatment increases average vessel length and
  • E E
  • FIG. 3A-D Nano-VEGF treatment does not alter Ataxin-l levels in SCA1 mice.
  • A Western blot of Ataxin-l mutant and wildtype shows protein levels were not affected by two weeks of VEGF treatment.
  • ANOVA analysis of variance
  • FIG. 4A-C Nano-VEGF treatment increases firing in Purkinje cells from SCA1 mice.
  • A Representative cell-attached recordings of spontaneous firing in Purkinje cells of l2-week-old mice treated with either vehicle (aCSF) or nano-VEGF. The PCs from VEGF treated mice (red trace) fired at higher frequencies than vehicle-treated mice (black trace).
  • B Representative probability distribution of inter-spike intervals in PC from vehicle-treated and VEGF-treated mice.
  • FIG. 5 VEGF levels are suppressed in SCA1 patients.
  • FIG. 6A-C VEGF effects in wild-type mice.
  • A Rotarod data for 26-week-old male WT mice treated with aCSF and VEGF shows overall no significant difference between the two groups over time.
  • B VEGF treatment of wild-type mice lengthened PC dendrites.
  • C rVEGF and nano-VEGF did not upregulate phosphorylation of VEGFR2 in WT mice, suggesting that there is a ceiling effect.
  • the term“comprise” and linguistic variations thereof denote the presence of recited feature(s), element(s), method step(s), etc. without the exclusion of the presence of additional feature(s), element(s), method step(s), etc.
  • the term “consisting of’ and linguistic variations thereof denotes the presence of recited feature(s), element(s), method step(s), etc. and excludes any unrecited feature(s), element(s), method step(s), etc., except for ordinarily-associated impurities.
  • the phrase“consisting essentially of’ denotes the recited feature(s), element(s), method step(s), etc. and any additional feature(s), element(s), method step(s), etc.
  • compositions, system, or method that do not materially affect the basic nature of the composition, system, or method.
  • Many embodiments herein are described using open “comprising” language. Such embodiments encompass multiple closed“consisting of’ and/or“consisting essentially of’ embodiments, which may alternatively be claimed or described using such language.
  • amino acid refers to natural amino acids, unnatural amino acids, and amino acid analogs, all in their D and L stereoisomers, unless otherwise indicated, if their structures allow such stereoisomeric forms.
  • Natural amino acids include alanine (Ala or A), arginine (Arg or R), asparagine (Asn or N), aspartic acid (Asp or D), cysteine (Cys or C), glutamine (Gln or Q), glutamic acid (Glu or E), glycine (Gly or G), histidine (His or H), isoleucine (Ile or I), leucine (Leu or L), Lysine (Lys or K), methionine (Met or M), phenylalanine (Phe or F), proline (Pro or P), serine (Ser or S), threonine (Thr or T), tryptophan (Trp or W), tyrosine (Tyr or Y) and valine (Val or V).
  • Unnatural amino acids include, but are not limited to, azetidinecarboxylic acid, 2- aminoadipic acid, 3-aminoadipic acid, beta-alanine, naphthylalanine (“naph”),
  • amino acid analog refers to a natural or unnatural amino acid where one or more of the C-terminal carboxy group, the N-terminal amino group and side-chain bioactive group has been chemically blocked, reversibly or irreversibly, or otherwise modified to another bioactive group.
  • aspartic acid-(beta-methyl ester) is an amino acid analog of aspartic acid
  • N-ethylglycine is an amino acid analog of glycine
  • alanine carboxamide is an amino acid analog of alanine.
  • amino acid analogs include methionine sulfoxide, methionine sulfone, S-(carboxymethyl)-cysteine, S-(carboxymethyl)- cysteine sulfoxide and S-(carboxymethyl)-cysteine sulfone.
  • peptide refers an oligomer to short polymer of amino acids linked together by peptide bonds. In contrast to other amino acid polymers (e.g., proteins, polypeptides, etc.), peptides are of about 50 amino acids or less in length.
  • a peptide may comprise natural amino acids, non-natural amino acids, amino acid analogs, and/or modified amino acids.
  • a peptide may be a subsequence of naturally occurring protein or a non-natural (artificial) sequence.
  • an artificial peptide, peptoid, or nucleic acid is one comprising a non-natural sequence (e.g., a peptide without 100% identity with a naturally-occurring protein or a fragment thereof).
  • a“conservative” amino acid substitution refers to the substitution of an amino acid in a peptide or polypeptide with another amino acid having similar chemical properties, such as size or charge.
  • each of the following eight groups contains amino acids that are conservative substitutions for one another:
  • Naturally occurring residues may be divided into classes based on common side chain properties, for example: polar positive (or basic) (histidine (H), lysine (K), and arginine (R)); polar negative (or acidic) (aspartic acid (D), glutamic acid (E)); polar neutral (serine (S), threonine (T), asparagine (N), glutamine (Q)); non-polar aliphatic (alanine (A), valine (V), leucine (L), isoleucine (I), methionine (M)); non-polar aromatic (phenylalanine (F), tyrosine (Y), tryptophan (W)); proline and glycine; and cysteine.
  • a“semi conservative” amino acid substitution refers to the substitution of an amino acid in a peptide or polypeptide with another amino acid within the same class.
  • a conservative or semi conservative amino acid substitution may also encompass non-naturally occurring amino acid residues that have similar chemical properties to the natural residue. These non-natural residues are typically incorporated by chemical peptide synthesis rather than by synthesis in biological systems. These include, but are not limited to, peptidomimetics and other reversed or inverted forms of amino acid moieties. Embodiments herein may, in some embodiments, be limited to natural amino acids, non-natural amino acids, and/or amino acid analogs.
  • Non-conservative substitutions may involve the exchange of a member of one class for a member from another class.
  • sequence identity refers to the degree of which two polymer sequences (e.g., peptide, polypeptide, nucleic acid, etc.) have the same sequential composition of monomer subunits.
  • sequence similarity refers to the degree with which two polymer sequences (e.g., peptide, polypeptide, nucleic acid, etc.) differ only by conservative and/or semi-conservative amino acid substitutions.
  • The“percent sequence identity” is calculated by: (1) comparing two optimally aligned sequences over a window of comparison (e.g., the length of the longer sequence, the length of the shorter sequence, a specified window, etc.), (2) determining the number of positions containing identical (or similar) monomers (e.g., same amino acids occurs in both sequences, similar amino acid occurs in both sequences) to yield the number of matched positions, (3) dividing the number of matched positions by the total number of positions in the comparison window (e.g., the length of the longer sequence, the length of the shorter sequence, a specified window), and (4) multiplying the result by 100 to yield the percent sequence identity or percent sequence similarity.
  • a window of comparison e.g., the length of the longer sequence, the length of the shorter sequence, a specified window, etc.
  • peptides A and B are both 20 amino acids in length and have identical amino acids at all but 1 position, then peptide A and peptide B have 95% sequence identity. If the amino acids at the non-identical position shared the same biophysical characteristics (e.g., both were acidic), then peptide A and peptide B would have 100% sequence similarity.
  • peptide C is 20 amino acids in length and peptide D is 15 amino acids in length, and 14 out of 15 amino acids in peptide D are identical to those of a portion of peptide C, then peptides C and D have 70% sequence identity, but peptide D has 93.3% sequence identity to an optimal comparison window of peptide C. For the purpose of calculating“percent sequence identity” (or“percent sequence similarity”) herein, any gaps in aligned sequences are treated as mismatches at that position.
  • any polypeptides described herein as having a particular percent sequence identity or similarity (e.g., at least 70%) with a reference sequence ID number may also be expressed as having a maximum number of substitutions (or terminal deletions) with respect to that reference sequence.
  • a sequence having at least Y% sequence identity (e.g., 90%) with SEQ ID NO:Z e.g., 100 amino acids
  • SEQ ID NO:Z e.g., 100 amino acids
  • X substitutions e.g., 10
  • nanofiber refers to an elongated or threadlike filament (e.g., having a significantly greater length dimension that width or diameter) with a diameter typically less than 100 nanometers.
  • the term“supramolecular” refers to the non-covalent interactions between molecules (e.g., polymers, marcomolecules, etc.) and the multicomponent assemblies, complexes, systems, and/or fibers that form as a result.
  • the terms“self-assemble” and“self-assembly” refer to formation of a discrete, non-random, aggregate structure from component parts; said assembly occurring spontaneously through random movements of the components (e.g. molecules) due only to the inherent chemical or structural properties and attractive forces of those components.
  • peptide amphiphile refers to a molecule that, at a minimum, includes a non-peptide lipophilic (hydrophobic) segment, a structural peptide segment and/or charged peptide segment (often both), and optionally a bioactive segment (e.g., linker segment, bioactive segment, etc.).
  • the peptide amphiphile may express a net charge at physiological pH, either a net positive or negative net charge, or may be zwitterionic (i.e., carrying both positive and negative charges).
  • Certain peptide amphiphiles consist of or comprise: (1) a hydrophobic, non-peptide segment (e.g., comprising an acyl group of six or more carbons), (2) a structural peptide segment (e.g., b-sheet forming); (3) a charged peptide segment, and (4) a bioactive segment (e.g., linker segment).
  • a hydrophobic, non-peptide segment e.g., comprising an acyl group of six or more carbons
  • a structural peptide segment e.g., b-sheet forming
  • a charged peptide segment e.g., linker segment
  • lipophilic moiety or “hydrophobic moiety” refers to the moiety (e.g., an acyl, ether, sulfonamide, or
  • a linear acyl chain is the lipophilic group (saturated or unsaturated carbons), palmitic acid.
  • other lipophilic groups may be used in place of the acyl chain such as steroids, phospholipids and fluorocarbons.
  • structural peptide refers to a portion of a peptide amphiphile, typically disposed between the hydrophobic segment and the charged peptide segment.
  • the structural peptide is generally composed of three to ten amino acid residues with non-polar, uncharged side chains (e.g., His (H), Val (V), Ile (I), Leu (L), Ala (A), Phe (F)) selected for their propensity to form hydrogen bonds or other stabilizing interactions (e.g., hydrophobic interactions, van der Waals' interactions, etc.) with structural segments of adjacent structural segments.
  • nanofibers of peptide amphiphiles having structural peptide segments display linear or 2D structure when examined by microscopy and/or a-helix and/or b-sheet character when examined by circular dichroism (CD).
  • beta ( )-sheet-forming peptide segment refers to a structural peptide segment that has a propensity to display b-sheet-like character (e.g., when analyzed by CD).
  • amino acids in a beta ⁇ )-sheet-forming peptide segment are selected for their propensity to form a beta-sheet secondary structure.
  • suitable amino acid residues selected from the twenty naturally occurring amino acids include Met (M), Val (V), Ile (I), Cys (C), Tyr (Y), Phe (F), Gln (Q), Leu (L), Thr (T), Ala (A), and Gly (G) (listed in order of their propensity to form beta sheets).
  • charged peptide segment refers to a portion of a peptide amphiphile that is rich (e.g., >50%, >75%, etc.) in charged amino acid residues, or amino acid residue that have a net positive or negative charge under physiologic conditions.
  • a charged peptide segment may be acidic (e.g., negatively charged), basic (e.g., positively charged), or zwitterionic (e.g., having both acidic and basic residues).
  • the terms“carboxy-rich peptide segment,”“acidic peptide segment,” and“negatively-charged peptide segment” refer to a peptide sequence of a peptide amphiphile that comprises one or more amino acid residues that have side chains displaying carboxylic acid side chains (e.g., Glu (E), Asp (D), or non-natural amino acids).
  • a carboxy- rich peptide segment may optionally contain one or more additional (e.g., non-acidic) amino acid residues.
  • Non-natural amino acid residues, or peptidomimetics with acidic side chains could be used, as will be evident to one ordinarily skilled in the art. There may be from about 2 to about 7 amino acids, and or about 3 or 4 amino acids in this segment.
  • amino-rich peptide segment refers to a peptide sequence of a peptide amphiphile that comprises one or more amino acid residues that have side chains displaying positively- charged acid side chains (e.g., Arg (R), Lys (K), His (H), or non-natural amino acids, or peptidomimetics).
  • a basic peptide segment may optionally contain one or more additional (e.g., non-basic) amino acid residues.
  • Non-natural amino acid residues with basic side chains could be used, as will be evident to one ordinarily skilled in the art. There may be from about 2 to about 7 amino acids, and or about 3 or 4 amino acids in this segment.
  • bioactive peptide refers to amino acid sequences that mediate the action of sequences, molecules, or supramolecular complexes associated therewith.
  • Peptide amphiphiles and structures (e.g., nanofibers) bearing bioactive peptides (e.g., a TF-targeting sequence, etc.) exhibits the functionality of the bioactive peptide.
  • biocompatible refers to materials and agents that are not toxic to cells or organisms.
  • a substance is considered to be “biocompatible” if its addition to cells in vitro results in less than or equal to approximately 10% cell death, usually less than 5%, more usually less than 1%.
  • biodegradable as used to describe the polymers, hydrogels, and/or wound dressings herein refers to compositions degraded or otherwise“broken down” under exposure to physiological conditions.
  • a biodegradable substance is a broken down by cellular machinery, enzymatic degradation, chemical processes, hydrolysis, etc.
  • a wound dressing or coating comprises hydrolyzable ester linkages that provide the biodegradability.
  • physiological conditions relates to the range of chemical (e.g., pH, ionic strength) and biochemical (e.g., enzyme concentrations) conditions likely to be encountered in the intracellular and extracellular fluids of tissues.
  • chemical e.g., pH, ionic strength
  • biochemical e.g., enzyme concentrations
  • the terms“treat,”“treatment,” and“treating” refer to reducing the amount or severity of a particular condition, disease state (e.g., SCA1), or symptoms thereof, in a subject presently experiencing or afflicted with the condition or disease state. The terms do not necessarily indicate complete treatment (e.g., total elimination of the condition, disease, or symptoms thereof).
  • Treatment encompasses any administration or application of a therapeutic or technique for a disease (e.g., in a mammal, including a human), and includes inhibiting the disease, arresting its development, relieving the disease, causing regression, or restoring or repairing a lost, missing, or defective function; or stimulating an inefficient process.
  • the terms“prevent,”“prevention,” and preventing” refer to reducing the likelihood of a particular condition or disease state (e.g., SCA1) from occurring in a subject not presently experiencing or afflicted with the condition or disease state.
  • the terms do not necessarily indicate complete or absolute prevention.
  • “preventing SCA1” refers to reducing the likelihood of SCA1 occurring in a subject not presently experiencing or diagnosed with SCAT
  • a composition or method need only reduce the likelihood of SCA1, not completely block any possibility thereof.
  • Prevention encompasses any administration or application of a therapeutic or technique to reduce the likelihood of a disease developing (e.g., in a mammal, including a human). Such a likelihood may be assessed for a population or for an individual.
  • the terms“co-administration” and“co-administering” refer to the administration of at least two agent(s) or therapies to a subject (e.g., a VEGF PA nanofiber and one or more therapeutic agents).
  • the co-administration of two or more agents or therapies is concurrent.
  • a first agent/therapy is administered prior to a second agent/therapy.
  • the appropriate dosage for co-administration can be readily determined by one skilled in the art.
  • the respective agents or therapies are administered at lower dosages than appropriate for their administration alone.
  • co-administration is especially desirable in embodiments where the co administration of the agents or therapies lowers the requisite dosage of a potentially harmful (e.g., toxic) agent(s), and/or when co-administration of two or more agents results in sensitization of a subject to beneficial effects of one of the agents via co-administration of the other agent.
  • a potentially harmful agent e.g., toxic
  • PAs self-assembling peptide amphiphiles
  • PAs comprising a bioactive vascular endothelial growth factor (VEGF) peptide, nanofibers displaying VEGF PAs, and methods of treatment or prevention of the polyglutamine disease Spinocerebellar Ataxia Type 1 (SCA1) and other neurodegenerative diseases therewith.
  • VEGF peptide delivery platform is provided in which the mechanical properties of the nanofiber material are tunable by altering the ratio of bioactive PA to structural PAs (e.g., acidic or basic PAs lacking a bioactive epitope).
  • Nano-VEGF activates the VEGF receptor (VEGFR2) and improves the SCA1 phenotype on both behavioral and pathological assays.
  • Nano-VEGF outperformed recombinant VEGF in several respects, such as improving the levels of capillary proteins and microvascular health.
  • VEGF VEGF receptor
  • VEGF vascular endothelial growth factor
  • VEGF vascular endothelial growth factor
  • Generating functional VEGF from bacterial and other expression systems is not only costly but inefficient, with considerable variations from one batch to the next (Storkebaum et al, 2005; herein incorporated by reference in its entirety). Beyond these practical limitations, recombinant VEGF has poor pharmacokinetics, with a very short half-life (Thome and Frey, 2001; Storkebaum et al, 2005; herein incorporated by reference in their entireties). Nano- VEGF, on the other hand, is stable (detectable in an ischemic limb model up to 4 weeks post injection) and forms small filamentous structures that break apart slowly, providing a slow- release formulation.
  • VEGF mimetic peptides on the surface is thought to promote receptor dimerization and sustained activation (Webber et al, 2011; herein incorporated by reference in its entirety). These parameters, contribute to nano- VEGF performing so favorably compared to recombinant VEGF.
  • VEGF peptide itself is designed to engage with VEGFR1 and 2 (Wiesmann et al, 1997; Diana et al, 2008; herein incorporated by reference in their entireties). While it is not the only region of VEGF that has been shown to activate these receptors, this peptide has been the best characterized in functional models. It is contemplated that two other domains (VEGF residues 61-66 and residues 79-93) also engage VEGF receptors.
  • peptide amphiphiles incorporating these sequences with that of the nano-VEGF peptide tested herein, and/or nanofibers comprising PAs displaying different VEGF peptides (e.g., 17-25, 61-66, and/or 79-93).
  • Intracerebroventricular (ICV) delivery of nano-VEGF was conducted to avoid systemic effects (e.g., promoting tumorigenesis (Carmeliet, 2005; herein incorporated by reference in its entirety), hypotension (Yang et al, 2002; herein incorporated by reference in its entirety), or coagulation disorders (Verheul et al., 2010; herein incorporated by reference in its entirety)).
  • ICV intraparenchymal delivery
  • VEGF by ICV enters the cerebrospinal fluid and bathes the brain, all circuits and cell types are being targeted.
  • FDA-approved battery-operated pumps that deliver drugs via a subcutaneously placed catheter into the CSF for long-term drug delivery are available, which obviate the need for repeated
  • VEGF levels are abnormally suppressed in several neurodegenerative diseases, including ALS, spinobulbar muscular atrophy (SBMA), Parkinson’s, and Alzheimer’s disease.
  • SBMA spinobulbar muscular atrophy
  • Parkinson Parkinson’s
  • Alzheimer Alzheimer’s disease.
  • nano-VEGF is provided for the
  • the peptide amphiphile molecules and compositions of the embodiments described herein are synthesized using preparatory techniques well-known to those skilled in the art, preferably, by standard solid-phase peptide synthesis, with the addition of a fatty acid in place of a standard amino acid at the N-terminus (or C-terminus) of the peptide, in order to create the lipophilic segment (although in some embodiments, alignment of nanofibers is performed via techniques not previously disclosed or used in the art (e.g., extrusion through a mesh screen).
  • Synthesis typically starts from the C-terminus, to which amino acids are sequentially added using either a Rink amide resin (resulting in an - NH2 group at the C-terminus of the peptide after cleavage from the resin), or a Wang resin (resulting in an—OH group at the C-terminus).
  • Rink amide resin resulting in an - NH2 group at the C-terminus of the peptide after cleavage from the resin
  • Wang resin resulting in an—OH group at the C-terminus.
  • some embodiments described herein encompass peptide amphiphiles having a C-terminal moiety that may be selected from the group consisting of— H, -OH, --COOH,— CONH2, and— NH2.
  • peptide amphiphiles comprise a hydrophobic (non-peptide) segment linked to a peptide.
  • the peptide comprises a structural segment (e.g., hydrogen-bond-forming segment, beta-sheet-forming segment, etc.), and a charged segment (e.g., acidic segment, basic segment, zwitterionic segment, etc.).
  • the peptide further comprises linker or spacer segments for adding solubility, flexibility, distance between segments, etc.
  • peptide amphiphiles comprise a spacer segment (e.g., peptide and/or non-peptide spacer) at the opposite terminus of the peptide from the hydrophobic segment.
  • the spacer segment comprises peptide and/or non-peptide elements.
  • the spacer segment comprises one or more bioactive groups (e.g., alkene, alkyne, azide, thiol, etc.).
  • various segments may be connected by linker segments (e.g., peptide (e.g., GG) or non-peptide (e.g., alkyl, OEG, PEG, etc.) linkers).
  • the lipophilic or hydrophobic segment is typically incorporated at the N- or C- terminus of the peptide after the last amino acid coupling, and is composed of a fatty acid or other acid that is linked to the N- or C-terminal amino acid through an acyl bond.
  • PA molecules self-assemble (e.g., into cylindrical micelles (a.k.a., nanofibers)) that bury the lipophilic segment in their core and display the bioactive peptide on the surface.
  • the structural peptide undergoes intermolecular hydrogen bonding to form beta sheets that orient parallel to the long axis of the micelle.
  • compositions described herein comprise PA building blocks that in turn comprise a hydrophobic segment and a peptide segment.
  • a hydrophobic (e.g., hydrocarbon and/or alkyl/alkenyl/alkynyl tail, or steroid such as cholesterol) segment of sufficient length e.g., 2 carbons, 3 carbons, 4 carbons, 5 carbons, 6 carbons, 7 carbons, 8 carbons, 9 carbons, 10 carbons, 11 carbons, 12 carbons, 13 carbons, 14 carbons, 15 carbons, 16 carbons, 17 carbons, 18 carbons, 19 carbons, 20 carbons, 21 carbons, 22 carbons, 23 carbons, 24 carbons, 25 carbons, 26 carbons, 27 carbons, 28 carbons, 29 carbons, 30 carbons or more , or any ranges there between.) is covalently coupled to peptide segment (e.g., a peptide comprising a segment having a preference for beta-strand conformations or other supramole
  • a plurality of such PAs will self-assemble in water (or aqueous solution) into a nanostructure (e.g., nanofiber).
  • the relative lengths of the peptide segment and hydrophobic segment result in differing PA molecular shape and nanostructural architecture.
  • a broader peptide segment and narrower hydrophobic segment results in a generally conical molecular shape that has an effect on the assembly of PAs (See, e.g., J. N. Israelachvib Intermolecular and surface forces; 2nd ed.; Academic: London San Diego, 1992; herein incorporated by reference in its entirety).
  • Other molecular shapes have similar effects on assembly and nanostructural architecture.
  • the pH of the solution may be changed (raised or lowered) or multivalent ions, such as calcium, or charged polymers or other macromolecules may be added to the solution.
  • the hydrophobic segment is a non-peptide segment (e.g., alkyl/alkenyl/alkynyl group).
  • the hydrophobic segment comprises an alkyl chain (e.g., saturated) of 4-25 carbons (e.g., 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,
  • the hydrophobic segment comprises an acyl/ether chain (e.g., saturated) of 2- 30 carbons (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30).
  • PAs comprise one or more peptide segments.
  • Peptide segment may comprise natural amino acids, modified amino acids, unnatural amino acids, amino acid analogs, peptidomimetics, or combinations thereof.
  • peptide segment comprise at least 50% sequence identity or similarity (e.g., conservative or semi
  • peptide amphiphiles comprise a charged peptide segment.
  • the charged segment may be acidic, basic, or zwitterionic.
  • peptide amphiphiles comprise an acidic peptide segment.
  • the acidic peptide comprises one or more (e.g., 1, 2, 3, 4, 5,
  • an acidic peptide segment comprises up to 7 residues in length and comprises at least 50% acidic residues.
  • an acidic peptide segment comprises (Xa)i-7, wherein each Xa is independently D or E.
  • an acidic peptide segment comprises EE.
  • peptide amphiphiles comprise a basic peptide segment.
  • the acidic peptide comprises one or more (e.g., 1, 2, 3, 4, 5,
  • an acidic peptide segment comprises (Xb)i-7, wherein each Xb is independently R, H, and/or K.
  • peptide amphiphiles comprises a structural and/or beta-sheet- forming segment.
  • the structural segment is rich in H, I, L, F, V, and A residues.
  • the structural and/or beta-sheet-forming segment comprises an alanine- and valine-rich peptide segment (e.g., AAVV (SEQ ID NO: 3), AAAVVV (SEQ ID NO:2), or other combinations of V and A residues, etc.).
  • the structural and/or beta sheet peptide comprises 4 or more consecutive A and/or V residues, or conservative or semi-conservative substitutions thereto.
  • the structural and/or beta-sheet forming peptide segment comprises 4 or more consecutive non-polar aliphatic residues (e.g., alanine (A), valine (V), leucine (L), isoleucine (I), methionine (M)).
  • non-polar aliphatic residues e.g., alanine (A), valine (V), leucine (L), isoleucine (I), methionine (M)
  • the structural and/or beta-sheet forming peptide segment comprises 2- 16 amino acids in length and comprises 4 or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,
  • peptide amphiphiles comprise a non-peptide spacer or linker segment.
  • the non-peptide spacer or linker segment is located at the opposite terminus of the peptide from the hydrophobic segment.
  • the spacer or linker segment provides the attachment site for a bioactive group.
  • the spacer or linker segment provides a reactive group (e.g., alkene, alkyne, azide, thiol, maleimide etc.) for functionalization of the PA.
  • the spacer or linker is a substantially linear chain of CH2, O, (CH 2 ) 2 0.
  • a spacer or linker further comprises additional bioactive groups, substituents, branches, etc.
  • Suitable peptide amphiphiles for use in the materials herein, as well as methods of preparation of PAs and realated materials, amino acid sequences for use in PAs, and materials that find use with PAs, are described in the following patents: U.S. Pat. No. 9,044,514; U.S. Pat. No. 9,040,626; U.S. Pat. No. 9,011,914; U.S. Pat. No. 8,772,228; U.S. Pat. No.
  • the characteristics (e.g., shape, rigidity, hydrophilicity, etc.) of a PA supramolecular structure depend upon the identity of the components of a peptide amphiphile (e.g., lipophilic segment, acidic segment, structural segment, bioactive segment, etc.).
  • a peptide amphiphile e.g., lipophilic segment, acidic segment, structural segment, bioactive segment, etc.
  • nanofibers, nanospheres, intermediate shapes, and other supramolecular structures are achieved by adjusting the identity of the PA component parts.
  • characteristics of supramolecular nanostructures of PAs are altered by post-assembly manipulation (e.g., heating/cooling, stretching, etc.).
  • a peptide amphiphile comprises: (a) a hydrophobic tail comprising an alkyl chain of 8-24 carbons; (b) a structural segment (e.g., comprising VVAA (SEQ ID NO: 4)); and (c) a charged segment (e.g., comprising KK, EE, etc.).
  • a structural segment e.g., comprising VVAA (SEQ ID NO: 4)
  • a charged segment e.g., comprising KK, EE, etc.
  • peptide amphiphiles comprise a bioactive moiety (e.g., VEGF peptide).
  • a bioactive moiety is the most C-terminal or N-terminal segment of the PA.
  • the bioactive moiety is attached to the end of the charged segment.
  • the bioactive moiety is exposed on the surface of an assembled PA structure (e.g., nanofiber).
  • a bioactive moiety is typically a peptide (e.g., gro VEGF peptide, etc.), but is not limited thereto.
  • a bioactive moiety is a peptide sequence that binds a peptide or polypeptide of interests, for example, a growth factor.
  • a VEGF peptide is provided as a PA bioactive moiety.
  • such VEGF peptide comprise at least 70% (e.g., 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or ranges therebetween) sequence identity with SEQ ID NO: 1.
  • a VEGF peptide is SEQ ID NO: 1.
  • nanofibers comprising bioactive PAs displaying one or more of a peptide comprising at least 70% (e.g., 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or ranges therebetween) sequence identity with one of SEQ ID NO: 1; a peptide comprising at least 70% (e.g., 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or ranges therebetween) sequence identity with residues 61-66 of VEGF; and/or a peptide comprising at least 70% (e.g., 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or ranges therebetween) sequence identity with residues 79-93 of VEGF.
  • a bioactive peptide comprises conservative or semi-conservative substitutions relative to one of SEQ ID NO: 1, residues 61-66 of VEGF, and/or residues 79-93 of VEGF.
  • a peptide amphiphile comprises: (a) a hydrophobic tail comprising an alkyl chain of 8-24 carbons; (b) a structural segment (e.g., comprising VVAA (SEQ ID NO: 4), AAVV (SEQ ID NO: 3), VA, AV, etc.); (c) a charged segment (e.g., comprising KK, EE, EK, KE, etc.), and a bioactive peptide (e.g., VEGF peptide).
  • a PA further comprises an attachment segment or residue (e.g., K) for attachment of the hydrophobic tail to the peptide potion of the PA.
  • the hydrophobic tail is attached to a lysine side chain.
  • a peptide amphiphile comprises (e.g., from C-terminus to N- terminus or fromN-terminus to C-terminus): bioactive peptide (e.g., VEGF peptide) - charged segment (e.g., comprising KK, EE, EK, KE, etc.) - structural segment (e.g., comprising VVAA (SEQ ID NO: 4), AAVV (SEQ ID NO: 3), VA, AV, etc.) - hydrophobic tail (e.g., comprising an alkyl chain of 8-24 carbons).
  • bioactive peptide e.g., VEGF peptide
  • charged segment e.g., comprising KK, EE, EK, KE, etc.
  • structural segment e.g., comprising VVAA (SEQ ID NO: 4), AAVV (SEQ ID NO: 3), VA, AV, etc.
  • hydrophobic tail e.g., comprising an alky
  • a peptide amphiphile comprises (e.g., from C-terminus to N- terminus or fromN-terminus to C-terminus): bioactive peptide (e.g., VEGF peptide) - charged segment (e.g., comprising KK, EE, EK, KE, etc.) - structural segment (e.g., comprising VVAA (SEQ ID NO: 4), AAVV (SEQ ID NO: 3) folk AV, etc.) - atachment segment or peptide (e.g., K) - hydrophobic tail (e.g., comprising an alkyl chain of 8-24 carbons).
  • bioactive peptide e.g., VEGF peptide
  • charged segment e.g., comprising KK, EE, EK, KE, etc.
  • structural segment e.g., comprising VVAA (SEQ ID NO: 4), AAVV (SEQ ID NO: 3) groove AV, etc.
  • a peptide amphiphile comprises (e.g., from C-terminus to N- terminus or fromN-terminus to C-terminus): bioactive peptide (e.g., growth factor or GF- targeting peptide) - KKAAVV(K) (SEQ ID NO: 5) - hydrophobic tail (e.g., comprising an alkyl chain of 8-24 carbons). In some embodiments, the hydrophobic tail is atached to the (K) sidechain.
  • bioactive peptide e.g., growth factor or GF- targeting peptide
  • KKAAVV(K) SEQ ID NO: 5
  • hydrophobic tail e.g., comprising an alkyl chain of 8-24 carbons.
  • the hydrophobic tail is atached to the (K) sidechain.
  • nanofibers and nanostructures assembled from the peptide amphiphiles described herein are provided herein.
  • a nanofiber is prepared by the self-assembly of the PAs described herein.
  • a nanofiber comprises or consists of PAs displaying a VEGF peptide.
  • the VEGF peptides are displayed on the surface of the nanofiber.
  • filler PAs are included in the nanofibers.
  • filler PAs are peptide amphiphiles, as described herein (e.g., structural segment, charged segment, hydrophobic segment, etc.), but lacking a bioactive moiety.
  • filler peptides are basic or acidic peptides lacking a bioactive moiety (e.g., V3A3K3, V3A3E3, etc.).
  • the filler PAs and VEGF PAs self- assemble into a nanofiber comprising both types of PAs.
  • nanostructures e.g., nanofibers assembled from the peptide amphiphiles described herein are provided.
  • filler peptides e.g., basic peptide, acidic peptides, etc.
  • filler peptides impart mechanical charateristics to a material comprising the PA nanofibers described herein.
  • a nanofiber assembled from 0-75% (mass%) bioactive VEGF PA and 25-100% (mass%) basic filler PA e.g., C16-VVVAAAKKK (SEQ ID NO: 6)
  • becomes a gel at basic pH conditions e.g., pH 8.5-11).
  • a nanofiber assembled from 75-100% (mass%) bioactive VEGF PA and 0-25% (mass%) basic filler PA e.g., Ci6- VVVAAAKKK (SEQ ID NO: 6)
  • basic filler PA e.g., Ci6- VVVAAAKKK (SEQ ID NO: 6)
  • a nanofiber assembled from 0-20% (mass%) bioactive VEGF PA and 80-100% (mass%) acidic filler PA e.g., C16-VVVAAAEEE (SEQ ID NO: 7)
  • becomes a gel at acidic pH conditions e.g., pH 1-5).
  • a nanofiber assembled from 20-80% (mass%) bioactive VEGF PA and 20-80% (mass%) acidic filler PA becomes a gel at neutral pH conditions (e.g., pH 5-8.5).
  • a nanofiber assembled from 80-100% (mass%) bioactive VEGF PA and 0-20% (mass%) acidic filler PA e.g., C16-VVVAAAEEE (SEQ ID NO: 7)
  • is a liquid at acidic pH conditions e.g., pH 1-5).
  • nanostructures are assembled from (1) PAs bearing a bioactive moiety (e.g., VEGF peptide) and (2) filler PAs (e.g., acidic or basic PAs not-labeled or not displaying a bioactive moiety, etc.).
  • PAs bearing a bioactive moiety e.g., VEGF peptide
  • filler PAs e.g., acidic or basic PAs not-labeled or not displaying a bioactive moiety, etc.
  • nanostructures e.g., nanofibers
  • nanostructures comprise 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50% 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 5%, 4%, 3%, 2%, or 1% (or any ranges there between) acidic filler PAs.
  • nanostructures e.g., nanofibers
  • nanofibers comprise 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50% 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 5%, 4%, 3%, 2%, or 1% (or any ranges there between) basic filler PAs.
  • the ratio of VEGF PA to acidic and/or basic PAs in a naofiber determines the mechanical characteristics (e.g., liquid or gel) of the nanofiber material and under what conditions the material will adopt various characteristics (e.g., gelling upon exposure to physiologic conditions, liquifying upon exposure to physiologic conditions, etc.).
  • Peptide amphiphile (PA) nanofiber solutions may comprise any suitable combination of PAs.
  • at least 0.05mg/mL e.g., O.lOmg/ml, 0. l5mg/ml, 0.20mg/ml, 0.25mg/ml, 0.30mg/ml, 0.35mg/ml, 0.40mg/ml, 0.45mg/ml, 0.50mg/ml, 0.60mg/ml, 0.70mg/ml, 0.80mg/ml, 0.90mg/ml, l.Omg/ml, or more, or ranges therebetween), of the solution is a filler PA (e.g., without a peptide epitope or other nanofiber surface displayed moiety).
  • a filler PA is a non-bioactive PA molecule having highly charged glutamic acid residues on the terminal end of the molecule (e.g., surface-displayed end). These negatively charged PAs allow for the gelation to take place between nanofibers via ionic crosslinks.
  • a filler PA is a non-bioactive PA molecule having highly charged lysine cid residues on the terminal end of the molecule (e.g., surface-displayed end). These positively charged PAs allow for the gelation to take place under basic conditions.
  • the filler PAs provide the ability to incorporate other bio-active PAs molecules into the nanofiber matrix while still ensuring the ability of the nanofibers solution to gel.
  • the solutions are annealed for increased viscosity and stronger gel mechanics.
  • These filler PAs have sequences are described in, for example, U.S. Pat. No. 8,772,228 (e.g., Ci6- VVVAAAEEE (SEQ ID NO: 7)), which is herein incorporated by reference in its entirety.
  • the PA nanofiber described herein exhibit a small cross- sectional diameter (e.g., ⁇ 25 nm, ⁇ 20 nm, ⁇ l5nm, about 10 nm, etc.). In some embodiments, the small cross-section of the nanofibers (-10 nm diameter) allows the fibers to permeate the brain parenchyma.
  • the PAs and nanofibers described herein find use in the treatment or prevention of neurodegenerative diseases, poly glutamine diseases, and in particular Spinocerebellar Ataxia Type 1 (SCA1).
  • SCA1 Spinocerebellar Ataxia Type 1
  • the PAs and nanofibers described herein find use in the treatment or prevention of neurodegenerative diseases, traumatic or mechanical injury to the central nervous system (CNS), spinal cord or peripheral nervous system (PNS), or other diseases or conditions of the nervous system.
  • diseases and conditions include, but are not limited to, Alzheimer's disease (AD), Parkinson's disease (PD),
  • Huntington's disease (HD), spinocerebellar ataxias, amyotrophic lateral sclerosis (ALS; Lou Gehrig's disease), diffuse Lewy body disease, chorea-acanthocytosis, primary lateral sclerosis, ocular diseases (ocular neuritis), chemotherapy -induced neuropathies (e.g., from vincristine, paclitaxel, bortezomib), diabetes-induced neuropathies and Friedreich's ataxia.
  • ALS amyotrophic lateral sclerosis
  • ocular diseases ocular neuritis
  • chemotherapy -induced neuropathies e.g., from vincristine, paclitaxel, bortezomib
  • diabetes-induced neuropathies e.g., from vincristine, paclitaxel, bortezomib
  • Friedreich's ataxia e.g., from vincristine, paclitaxel, bortezomib
  • VEGF PA nanofibers herein find use in the treatment of dementia and dementia-related diseases and conditions.
  • Dementias are diseases that include memory loss and additional intellectual impairment separate from memory.
  • the VEGF PA nanofibers herein are suitable for use in treating patients suffering from memory impairment in all forms of dementia. Dementias are classified according to their cause and include:
  • neurodegenerative dementias e.g., Alzheimer's, Parkinson's disease, Huntington's disease, Pick's disease
  • vascular e.g., infarcts, hemorrhage, cardiac disorders
  • mixed vascular and Alzheimer's bacterial meningitis, Creutzfeld-Jacob Disease, multiple sclerosis
  • traumatic e.g., subdural hematoma or traumatic brain injury
  • infectious e.g., HIV
  • toxic e.g., heavy metals, alcohol, some medications
  • metabolic e.g., vitamin B12 or folate deficiency
  • CNS hypoxia e.g., Cushing's disease
  • psychiatric e.g., depression and schizophrenia
  • hydrocephalus e.g., Alzheimer's, Parkinson's disease, Huntington's disease, Pick's disease
  • vascular e.g., infarcts, hemorrhage, cardiac disorders
  • the condition of memory impairment is manifested by impairment of the ability to leam new information and/or the inability to recall previously learned information.
  • the present invention includes methods for dealing with memory loss separate from dementia, including mild cognitive impairment (MCI) and age-related cognitive decline.
  • MCI mild cognitive impairment
  • the present invention includes methods of treatment for memory impairment as a result of disease.
  • Memory impairment is a primary symptom of dementia and can also be a symptom associated with such diseases as Alzheimer's disease, schizophrenia, Parkinson's disease, Huntington's disease, Pick's disease, Creutzfeld-Jakob disease, HIV, cardiovascular disease, and head trauma as well as age-related cognitive decline.
  • VEGF PA nanofibers herein are suitable for use in the treatment of memory impairment due to, for example, Alzheimer's disease, multiple sclerosis, amylolaterosclerosis (ALS), multiple systems atrophy (MSA), schizophrenia, Parkinson's disease, Huntington's disease, Pick's disease, Creutzfeld-Jakob disease, depression, aging, head trauma, stroke, spinal cord injury, CNS hypoxia, cerebral senility, diabetes associated cognitive impairment, memory deficits from early exposure of anesthetic agents, multiinfarct dementia and other neurological conditions including acute neuronal diseases, as well as HIV and cardiovascular diseases.
  • ALS amylolaterosclerosis
  • MSA multiple systems atrophy
  • schizophrenia Parkinson's disease
  • Huntington's disease Huntington's disease
  • Pick's disease Creutzfeld-Jakob disease
  • depression head trauma
  • stroke spinal cord injury
  • CNS hypoxia CNS hypoxia
  • cerebral senility diabetes associated cognitive impairment
  • memory deficits from early exposure of anesthetic agents multiinfarct dementia
  • the VEGF PA nanofibers herein are also suitable for use in the treatment of a class of disorders known as poly glutamine-repeat diseases. These diseases share a common pathogenic mutation.
  • the expansion of a CAG repeat, which encodes the amino acid glutamine, within the genome leads to production of a mutant protein having an expanded polyglutamine region.
  • Huntington's disease has been linked to a mutation of the protein huntingtin. In individuals who do not have Huntington's disease, huntingtin has a poly glutamine region containing about 8 to 31 glutamine residues. For individuals who have Huntington's disease, huntingtin has a poly glutamine region with over 37 glutamine residues.
  • DRPLA dentatorubral-pallidoluysian atrophy
  • DRPLA dentatorubral-pallidoluysian atrophy
  • DRPLA atrophin- 1
  • spinocerebellar ataxia type-l ataxin-l
  • spinocerebellar ataxia type-2 ataxin-2
  • spinocerebellar ataxia type-3 also called Machado-Joseph disease or MJD
  • MJD Machado-Joseph disease
  • spinocerebellar ataxia type-6 alpha la-voltage dependent calcium channel
  • spinocerebellar ataxia type-7 ataxin-7
  • spinal and bulbar muscular atrophy SBMA, also know as Kennedy disease.
  • compositions and methods herein find use in inducing regrowth of blood vessels for the treatment of vascular diseases like critical limb ischemia.
  • compositions and methods herein find use in the treatment or prevention of SCA1.
  • compositions and methods herein find use in activating cell signaling in vitro (e.g., as a cell culture manipulation tool).
  • the VEGF PA nanofiber compositions herein are formulated for delivery to a subject.
  • Suitable routes of administrating the pharmaceutical composition described herein include, without limitation: topical, subcutaneous, transdermal, intradermal, intralesional, intraarticular, intraperitoneal, intravesical, transmucosal, gingival, intradental, intracochlear, transtympanic, intraorgan, epidural, intrathecal, intramuscular, intravenous, intravascular, intraosseus, periocular, intratumoral, intracerebral,
  • VEGF PA nanofiber compositions are administered parenterally.
  • parenteral administration is by intrathecal administration, intracerebroventricular administration, or intraparenchymal administration.
  • VEGF PA nanofiber compositions herein can be administered as the sole active agent or in combination with other pharmaceutical agents such as other agents used in the treatment of neurological diseases and conditions, e.g., nicotinic alpha.-7 agonists, PDE4 inhibitors, other PDE10 inhibitors, calcium channel blockers, muscarinic ml and m2 modulators, adenosine receptor modulators, ampakines, NMDA-R modulators, mGluR modulators, dopamine modulators, serotonin modulators, canabinoid modulators, and cholinesterase inhibitors (e.g., donepezil, rivastigimine, and galanthanamine).
  • other pharmaceutical agents such as other agents used in the treatment of neurological diseases and conditions, e.g., nicotinic alpha.-7 agonists, PDE4 inhibitors, other PDE10 inhibitors, calcium channel blockers, muscarinic ml and m2 modulators, adenosine receptor modul
  • Drugs suitable in combination with the compounds of the present invention include, but are not limited to Clozaril, Zyprexa, Risperidone, Seroquel, Lithium, Zyprexa, Depakote, Levodopa, Parlodel, Permax, Mirapex, Tasmar, Contan, Kemadin, Artane, Cogentin, Reminyl, Akatinol, Neotropin, Eldepryl, Estrogen Cliquinol, Thioridazine, Haloperidol, Risperidone, Cognex, Aricept, and Exelon, Dilantin, Luminol, Tegretol, Depakote, Depakene, Zarontin, Neurontin, Barbita, Solfeton, Felbatol, Detrol, Ditropan XL, OxyContin, Betaseron, Avonex,
  • the Pathology Core Facility at Northwestern University performed immunohistochemistry for VEGF (Anti-VEGF ab39250; Abeam 1: 100, Cambridge, UK) on 5pm-thick human cerebellar tissues.
  • Optical density analysis was performed with background subtraction using ImageJ using color deconvolution method (Ruifrok and Johnston, 2001; herein incorporated by reference in its entirety).
  • mice were generated as described previously (Watase et al, 2002b; herein incorporated by reference in its entirety). These mice express a pathogenic poly glutamine tract of 154 repeats from one allele; the other allele expresses a normal mouse Atxnl with two repeats (normal human alleles range from 6 to 44 poly glutamine repeats) (Quan et al, 1995; Servadio et al, 1995; Goldfarb et al., 1996; herein incorporated by reference in their entireties). Initially generated on a C57BL/6J-l29SvEv mixed genetic background, the mice were backcrossed more than ten generations with C57BL/6J mice to eliminate any background effects.
  • mice were not separated by sex.
  • Nano-VEGF was engineered by linking to the N terminus of a VEGF peptide
  • the peptides were synthesized using 9-fluorenyl methoxy carbonyl (Fmoc) solid-phase peptide synthesis with rink amide 4- methylbenzhydrylamine resin (EMD Millipore). The synthesis was performed on a Liberty automated microwave peptide synthesizer (CEM Corp, Matthews, NC) at Northwestern’s Peptide Synthesis Core. Fmoc groups were removed with 30% 4-methylpiperidine and 0.1 M hydroxybenzotriazole (HOBt) in N,N-dimethylformamide (DMF) at 75°C for 3-4 min for each amino acid or palmitic acid (4 equiv.).
  • Fmoc 9-fluorenyl methoxy carbonyl
  • the peptides were coupled at 75 °C for 5-10 min using 4 equiv. of O-benzotriazole-N, N, N', N'-tetramethyluronium hexafluorophosphate (HBTU), and 8 equiv. of N,N-diisopropylethylamine (DIEA).
  • HBTU O-benzotriazole-N, N, N', N'-tetramethyluronium hexafluorophosphate
  • DIEA N,N-diisopropylethylamine
  • the newly synthesized amphiphilic peptides were cleaved from the resin with a 95:2.5:2.5 trifluoroacetic acid (TFA)/triisopropylsilane (TIPS)/water mixture for 3-4 h.
  • Rotary evaporation and precipitation in cold diethyl ether yielded the crude peptide mixture.
  • mice were treated for a two- week period starting at 24 weeks.
  • Nano-VEGF, mouse rVEGF (R&D systems, Minneapolis, MN) or artificial CSF (sham controls) were delivered using the intracerebroventricular route (into the right lateral ventricle with stereotaxic coordinates: A/P -0.5mm, M/L -l.lmm, D/V -2.5mm). using osmotic ALZET pumps; model#l002, Durect) (Cvetanovic et al, 2011; herein incorporated by reference in its entirety). Behavioral assays were performed at 26 weeks; while biochemical and histochemical analysis was performed at 27 weeks (after euthanasia). For electrophysiological experiments, we used 8-10 week-old mice for pump placement (with a similar two-week treatment regimen) since it is difficult to generate good electrophysiological traces from older mice.
  • Rotating Rod Assay 26-week-old mice (post-treatment) were placed on a rotating rod apparatus (Ugo Basile) that accelerates linearly from 4 to 40 rotations per minute over a 5- minute period. Mice were subjected to four trials per day for four consecutive days, each trial lasting to a maximum of ten minutes, with at least ten minutes of rest between each trial. The average performances for each day were plotted, and statistical differences between the different groups were analyzed using repeated measures two-way ANOVA (followed by Tukey’s honest significant difference post-hoc test for multiple comparisons). All statistical analyses here and elsewhere were performed using GraphPad Prism 6 software (GraphPad Software, La Jolla California USA). Data were considered significant for p ⁇ 0.05. Data are expressed as mean ⁇ SEM.
  • Gait Analysis Mice were placed on a transparent treadmill belt connected to a high speed digital video camera (Digigait ⁇ system; Mouse Specifics, Inc.). The speed of the treadmill was set to the minimum of 5 cm/s up to maximum tolerated speed (maxed out at 24 cm/s). The performance of mice on the treadmill at different speeds was recorded with digital video camera.
  • mice post treatment and post behavioral analysis were anesthetized by isoflurane inhalation and were perfused with ice-cold PBS, followed by 4%
  • Brain tissues were blocked and permeabilized with 5% Normal donkey serum in 1XTBS with 0.25% Triton-X 100 (TBST) for 2 hours at room temperature. Slices were then incubated with primary antibodies for 48- 72 hours in 4°C. After three washes in TBST, signals were detected using according secondary antibodies for 2 hours at RT. Brain slices were counterstained with DAPI
  • mice were studied per experimental group and performed analysis on 3- 5 sections per mouse. Images were taken as Z-stack images using the Leica TCS SP5 confocal microscope (Cvetanovic et al, 2011). To observe the morphology of Purkinje cells, calbindin staining (mouse anti-Calbindin-D-28K (C9848, Sigma, 1 :2000) was performed. To quantify dendritic length, the calbindin-stained PCs were traced and the length of the PC dendrites was measured from the proximal end of the soma to the distal end of the dendrites (using ImageJ l.46r Software; National Institutes of Health, Bethesda, MD, USA).
  • the vascular network was visualized by staining for Collagen IV (rabbit anti-Collagen IV (abl9808, Abeam, 1:400). Average vessel length and branching index was plotted and analyzed by assessing the variation in foreground and background pixel mass densities across images (using the open source AngioTool software (angiotool.nci.nih.gov; Zudaire et al, 2011; herein incorporated by reference in its entirety). Statistical differences for dendritic length and microvasculature parameters were compared using one-way ANOVA followed by Tukey’s post-hoc test.
  • brain sections were stained for ataxin-l (mouse anti-Ataxin-l 11NQ clone N76/3; NeuroMab, 1 : 1000) and counterstained with DAPI.
  • the number of cells with ataxin-l inclusions in each defined region of interest was normalized to the number of cells stained with DAPI only.
  • Statistical analysis was performed using unpaired t test. In all cases, the secondary antibodies were generated in donkey (donkey anti-mouse Alexa Fluor®647 and donkey anti-rabbit Alexa Fluor® 488; both at a dilution of 1 :500 Invitrogen,).
  • Proteins were extracted from cerebellar tissues using radioimmunoprecipitation assay (RIP A) buffer (50mM Tris-HCl, pH 8.0, l50mM NaCl, lmM EDTA, lOmM NaF, 0.5% Sodium deoxycholate, 1% Triton-X 100, 0.1% SDS, protease inhibitors (Sigma)), and resolved on 7.5% SDS-PAGE gel. Densitometric analysis was evaluated using ImageJ l.46r software (National Institutes of Health, Bethesda, MD, USA).
  • mouse anti-Ataxin-l 11NQ clone N76/3; NeuroMab, 1 :1000
  • rabbit anti-Claudin 5 34-1600, Thermo Fisher Scientific 1:200
  • rabbit anti-Occludin 71-1500, Thermo Fisher Scientific, 1 :200
  • mouse anti-ZO-l 33-9100, Thermo Fisher Scientific, 1: 100
  • mouse anti -beta Actin A2228, Sigma, 1:5000
  • the secondary antibodies used were anti-rabbit IgG (H+L) HRP (W4011, Promega, 1 :5000) or anti-mouse IgG (H+L) HRP (W4021, Promega, 1 :5000) secondary antibodies (visualized with ECL substrate (GE); film densitometric analysis was evaluated using ImageJ l.46r software (National Institutes of Health, Bethesda, MD, USA). Statistical differences were compared using one-way ANOVA followed by Tukey’s post-hoc test.
  • Phospho-VEGFR2 ELISA was performed using PathScan® Phospho-VEGFR2 (Tyl l75) Sandwich ELISA kit (7335, Cell Signaling), according to manufacturer’s instruction. Cerebellar lysates were prepared in 2X cell lysis buffer, and undiluted tissue lysates were incubated in 96-well plates coated with phosphor-VEGFR2 primary antibody overnight at 4°C. Wells were washed several times, and detection antibody was added to the well and incubated for 1 hr at 37C and with HRP -conjugated secondary antibody for 30 min at 37°C. Signals were detected with 3,3’,5,5’-Tetramethylbenzidine (TMB) substrate and sulfuric acid were added to stop the reaction. Absorbance was read at 450nm within 30 min after stopping the reaction.
  • TMB 3,3’,5,5’-Tetramethylbenzidine
  • mice were anesthetized with isoflurane and killed by decapitation.
  • the brains were removed from the skull and placed in warm (30-33°C) artificial cerebrospinal fluid (ACSF) containing (in mM): 125 NaCl, 25 NaHC03, 2.5 KC1, 1.25 NaH2P04, 0.5 CaCl2, 7 MgCl2, 75 sucrose and 25 glucose, equilibrated with 95% 02 and 5% C02 (pH 7.4).
  • 300 pm thick para-sagittal cerebellar slices were cut using a vibratome (Leica VT1200) and stored in the same solution at 34°C for 15 min and then at 20-22°C until used ( ⁇ 6 hours).
  • the slices were transferred to a chamber bathed in physiological ACSF (in mM: 125 NaCl, 25 NaHC03, 2.5 KC1, 1.25 NaH2P03, 25 glucose, 2.0 CaCl2 and 1 MgCl2, (equilibrated with 95% 02 and 5% C02) at 30-32°C.
  • Slices were visualized using an upright microscope (Scientifica) equipped with a 40X water-immersion objective (Olympus), oblique illumination, and video microscopy using a digital camera (Q- imaging). Pipettes for cell-attached recordings were pulled from borosilicate glass (Sutter) using a horizontal puller (P97, Sutter).
  • Tip resistances in working solution ranged from 3 to 6 MW when filled with physiological ACSF. Electrophysiological recordings were obtained using an Axopatch 200B amplifier, filtered at 20 KHz and acquired at 10 KHz. Data were obtained and visualized using pClamp9 software. Cells were recorded in the I-Clamp fast mode. For all recordings, fast synaptic transmission was blocked by 3 mM kynurenic acid (Sigma) and 0.1 mM picrotoxin (Abeam). Statistical analysis was performed using unpaired t test.
  • Nano-VEGF improves motor coordination in SCA1 mice with advanced disease
  • Cerebella of SCA1 knock-in mice show lower VEGF expression than wild-type (Cvetanovic et al., 2011; herein incorporated by reference in its entirety).
  • immunohistochemical analysis was performed on post-mortem cerebella obtained from SCA1 patients and age-matched controls. VEGF staining is clearly reduced in the cerebellar lobules of SCA1 patients (Fig. 5A and B).
  • SCA1 knock-in mice SCA1154Q/2Q
  • Nano-VEGF was tested against recombinant VEGF (rVEGF) on SCA1 mice at 24 weeks of age, which corresponds to advanced disease in humans. Any improvement in these mice would be a more stringent test of efficacy.
  • the compounds were delivered over a two- week period, rVEGF at a dose of 2.5 pg (dissolved in 100 pl of artificial CSF (aCSF) (Cvetanovic et al, 2011), and nano-VEGF at a dose of 20.0 pg [the bioequivalence of nano- VEGF was determined to be approximately eight times that of rVEGF based on functional studies in a mouse hind-limb ischemia model (Webber et al, 2011; herein incorporated by reference in its entirety). The effects of both forms of VEGF were then evaluated at the end of the delivery period by behavioral testing in the 26th week and neuropathological assessment in the 27th week (Fig. 1 A).
  • Gait analysis was performed using a videotaped assessment of gait on a transparent treadmill. SCA1 mice were barely able to walk on the moving treadmill even at low speeds (5 cm/s), but mice treated with nano-VEGF (or rVEGF) could walk and even run when the speed of the treadmill was increased to 17 cm/sec (tested up to 24 cm/s).
  • nano-VEGF or rVEGF
  • VEGF membrane- spanning VEGF receptor 2
  • VEGFR2 membrane- spanning VEGF receptor 2
  • VEGF receptor 2 a tyrosine kinase receptor formed of two monomers.
  • VEGF membrane- spanning VEGF receptor 2
  • the monomers dimerize to cause autophosphorylation of the intracellular tyrosine residues, which activates downstream signaling pathways that mediate its effects (Cross and Claesson-Welsh, 2001; herein incorporated by reference in its entirety).
  • ELISA showed a decrease in the level of VEGFR2 phosphorylation consistent with low levels of VEGF in SCA1 mice compared to controls.
  • mice treated with nano-VEGF showed a greater improvement in vessel length and branching index than mice treated with rVEGF (Fig. 2B-2D).
  • Western blot studies showed that reduced levels of capillary proteins (the tight junction markers ZO-l, Claudin-4, and Occludin (Chiba et al, 2008; Zhong et al., 2008; herein incorporated by reference in their entireties)) in SCA1 mice are restored with VEGF replenishment, and again nano-VEGF was more effective than rVEGF (Fig. 2E-2H).
  • Purkinje cells provide the only output of the cerebellar cortex and transmit the integrated activity of the cortex to the cerebellar nuclei.
  • PCs show less spontaneous firing not only because of their own pathology but also because of a decrease in climbing fiber (CF) excitatory postsynaptic currents (EPSCs) (Dell'Orco et al, 2015; Ruegsegger et al, 2016b; herein incorporated by reference in their entireties) and an increase in GABA-ergic inhibition from basket cells (Edamakanti et al, 2018; herein incorporated by reference in its entirety).
  • CF climbing fiber
  • EPCs excitatory postsynaptic currents
  • Nano-VEGF -treated mice showed significantly faster and more regular firing compared with non-treated SCA1 mice, in keeping with the behavioral improvement (Fig. 4A-C).
  • Nano-VEGF thus activates VEGF signaling receptors, improves pathology and enhances the motor ability of SCA1 mice.
  • the disease-modifying effects are quite remarkable, given that the treatment regimen was initiated only after neurodegeneration was well-established.
  • Vascular endothelial growth factor ameliorates the ataxic phenotype in a mouse model of spinocerebellar ataxia type 1. Nature medicine 2011; 17(11): 1445-7.
  • VEGF vascular endothelial growth factor
  • VEGF vascular endothelial growth factor
  • VEGF Vascular endothelial growth factor
  • Ruifrok AC Johnston DA. Quantification of histochemical staining by color deconvolution.
  • Thome RG Frey WH, 2nd. Delivery of neurotrophic factors to the central nervous system: pharmacokinetic considerations. Clinical pharmacokinetics 2001; 40(12): 907-46.
  • HDAC3 histone deacetylase HDAC3 is essential for Purkinje cell function, potentially complicating the use of HD AC inhibitors in SCAT Hum Mol Genet 2014; 23(14): 3733-45.
  • Verheul HM van Erp K
  • Homs MY Yoon GS
  • van der Groep P Rogers C, et al.

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Abstract

La présente invention concerne des amphiphiles peptidiques auto-assemblées (PA) comprenant un facteur de croissance de l'endothélium vasculaire (VEGF) bioactif et des méthodes de traitement ou de prévention de l'ataxie spinocérébelleuse de type 1 (SCA1) et d'autres maladies neurodégénératives associées. Dans des modes de réalisation particuliers, l'invention concerne une plate-forme d'administration de peptide VEGF dans laquelle les propriétés mécaniques du matériau de nanofibres sont ajustables par modification du rapport de PA bioactif à des PA structuraux (par exemple, PA acides ou basiques dépourvus d'épitope bioactif).
PCT/US2019/060466 2018-11-09 2019-11-08 Nanoparticules de vegf auto-assemblées Ceased WO2020097460A1 (fr)

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US20120294902A1 (en) * 2011-04-08 2012-11-22 Stupp Samuel I Peptide amphiphiles and methods to electrostatically control bioactivity of the ikvav peptide epitope
US20130101628A1 (en) * 2011-04-29 2013-04-25 Northwestern University Novel vegf mimetic peptide-based scaffolds for therapeutic angiogenesis and methods for their use

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US20120294902A1 (en) * 2011-04-08 2012-11-22 Stupp Samuel I Peptide amphiphiles and methods to electrostatically control bioactivity of the ikvav peptide epitope
US20130101628A1 (en) * 2011-04-29 2013-04-25 Northwestern University Novel vegf mimetic peptide-based scaffolds for therapeutic angiogenesis and methods for their use

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